Supplementary Information Molecular mechanism of male differentiation is conserved in the SRY-absent mammal, Tokudaia osimensis Tomofumi Otake1 & Asato Kuroiwa1, 2 1
Functional Genome Science Biosystems Science Course, Graduate School of Life Science,
Hokkaido University, Kita 10 Nishi 8, Kita-ku, Sapporo, Hokkaido 060-0810, Japan 2
Division of Reproductive and Developmental Biology, Department of Biological Sciences,
Faculty of Science, Hokkaido University, Kita 10 Nishi 8, Kita-ku, Sapporo, Hokkaido 060-0810, Japan Tel/Fax: +81-11-706-2752; E-mail:
[email protected]
Correspondence: A. Kuroiwa
Supplementary Figure Legends Figure S1. Sequence alignment of the mouse, rat, and T. osimensis AMH proximal promoter (A) Sequence alignment of the AMH proximal promoter. The SF1 BS, SOX BS, GATA4 BS, and WT1 BS are boxed with black, red, blue, and gray squares, respectively. +1 marks the transcriptional start site in the mouse. (B) Sequence alignment in the SOX9 proximal promoter. The ETS BS sequences (5’-GGAA/T-3’, 5’-T/ATCC-3’ in reverse) are denoted in gray. The number of the nucleotide corresponds to the T. osimensis sequence.
Figure S2. Mutagenesis of the T. osimensis AMH promoter
Nucleotide substitutions were introduced in the SF1 BS (R1, regulatory mutation-1), SOX BS (R2, regulatory mutation-2), and both SF1 BS and SOX BS in cis (R3, regulatory mutation-3). These mutations were reported in previous reports for mouse22, 27.
Figure S3. Expression of Sox9/SOX9, Sox8/SOX8, Sox10/SOX10 and Er71/ER71 in mice and T. osimensis The expression patterns of Sox9/SOX9, Sox8/SOX8, Sox10/SOX10 (A) and Er71/ER71 (B) in male and female mice and T. osimensis by RT-PCR. H, heart; Li, liver; K, kidney; S, spleen; B, brain; Lu, lung; T, testis; O, ovary. Actb/ACTB was used as the internal control. For the negative control, distilled sterile water (dH2O) was used as the template.
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